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( Gopal Prasad Ghimire ),( Niranjan Koirala ),( Jae Kyung Sohng ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.5
Genes encoding enzymes with sequence similarity to hopanoids biosynthetic enzymes of other organisms were cloned from the hopanoid (hop) gene cluster of Streptomyces peucetius ATCC 27952 and transformed into Streptomyces venezuelae YJ028. The cloned fragments contained four genes, all transcribed in one direction. These genes encode polypeptides that resemble polyprenyl diphosphate synthase (hopD), squalene-phytoene synthases (hopAB), and squalenehopene cyclase (hopE). These enzymes are sufficient for the formation of the pentacyclic triterpenoid lipid, hopene. The formation of hopene was verified by gas chromatography/ mass spectrometry.
Gopal Prasad Ghimire,오태진,류광경,송재경 한국분자세포생물학회 2008 Molecules and cells Vol.26 No.4
We identified a 1,134-bp putative type III polyketide synthase from the sequence analysis of Streptomyces peucetius ATCC 27952, named Sp-RppA, which is characterized as 1,3,6,8-tetrahydroxynaphthalene synthase and shares 33% identity with SCO1206 from S. coelicolor A3(2) and 32% identity with RppA from S. griseus. The 1,3,6,8-tetrahydrox-ynaphthalene synthase is known to catalyze the sequential decarboxylative condensation, intramolecular cyclization, and aromatization of an oligoketide derived from five units of malonyl-CoA to give 1,3,6,8-tetrahydroxynaphthalene, which spontaneously oxidizes to form 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin). In this study, we report the in vivo expression and in vitro synthesis of flaviolin from purified gene product (Sp-RppA).
Review : Advances in Biochemistry and Microbial Production of Squalene and Its Derivatives
( Gopal Prasad Ghimire ),( Nguyen Huy Thuan ),( Niranjan Koirala ),( Jae Kyung Sohng ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.3
Squalene is a linear triterpene formed via the MVA or MEP biosynthetic pathway and is widely distributed in bacteria, fungi, algae, plants, and animals. Metabolically, squalene is used not only as a precursor in the synthesis of complex secondary metabolites such as sterols, hormones, and vitamins, but also as a carbon source in aerobic and anaerobic fermentation in microorganisms. Owing to the increasing roles of squalene as an antioxidant, anticancer, and anti-inflammatory agent, the demand for this chemical is highly urgent. As a result, with the exception of traditional methods of the isolation of squalene from animals (shark liver oil) and plants, biotechnological methods using microorganisms as producers have afforded increased yield and productivity, but a reduction in progress. In this paper, we first review the biosynthetic routes of squalene and its typical derivatives, particularly the squalene synthase route. Second, typical biotechnological methods for the enhanced production of squalene using microbial cell factories are summarized and classified. Finally, the outline and discussion of the novel trend in the production of squalene with several updated events to 2015 are presented.
Ghimire, Gopal Prasad,Lee, Hei Chan,Sohng, Jae Kyung American Society for Microbiology 2009 Applied and environmental microbiology Vol.75 No.22
<B>ABSTRACT</B><P>Putative hopanoid genes from <I>Streptomyces peucetius</I> were introduced into <I>Escherichia coli</I> to improve the production of squalene, an industrially important compound. High expression of <I>hopA</I> and <I>hopB</I> (encoding squalene/phytoene synthases) together with <I>hopD</I> (encoding farnesyl diphosphate synthase) yielded 4.1 mg/liter of squalene. This level was elevated to 11.8 mg/liter when there was also increased expression of <I>dxs</I> and <I>idi</I>, <I>E. coli</I> genes encoding 1-deoxy-d-xylulose 5-phosphate synthase and isopentenyl diphosphate isomerase.</P>
Hybrid Breeding in Vegetables in Nepal
Tek Prasad Gotame,Ishwori Prasad Gautam,Surendra Lal Shrestha,Navin Gopal Pradhan 한국국제농업개발학회 2021 韓國國際農業開發學會誌 Vol.33 No.4
In Nepal, about 73% of the vegetable production area is estimated to be covered by hybrid varieties. Most of the hybrid seeds in vegetable crops are imported from abroad. Considering the demand for hybrid vegetable varieties, the National Horticulture Research Centre (NHRC), Khumaltar, Lalitpur has started hybrid breeding of vegetables using a conventional approach since the fiscal year 2003/04. With this effort, the tomato hybrid ‘Srijana’ was developed and registered in 2010. This variety was estimated to cover about 35% of the total tomato cultivated area. Two tomato hybrids, namely ‘Khumal Hybrid Tomato-2 (HRA14 × HRD7) and ‘Khumal Hybrid Tomato-3’ (HRA20 × HRD2), showed good performance and were released in the National Seed Board in 2021. It was found that the heterosis for the yield was 87% in ‘Khumal Hybrid Tomato-2’ and 95% in ‘Khumal Hybrid Tomato-3’ with a potential yield of 70 MT/ha and 60 MT/ha, respectively, under open field conditions. Similarly, a comparative study of 7 tomato hybrids at different research stations was carried out and it showed that F1 (HRA13 × HRD7) could be a potential hybrid for the western region with a potential productivity of 116.8 MT/ha under a protected cultivation system. Hybrid breeding of cucumber was also initiated in the fiscal year 2007/08. A total of 12 crosses in cucumber were performed and two hybrids F1 (HRDCUC004 x RDCUC001), 107 MT/ha and F1 (HRDCUC009 × HRDCUC003) 150 MT/ha were found promising. Hybrid breeding in brinjal, hot pepper and bitter gourd has also been started. Furthermore, key problems and way forward to strengthen hybrid vegetable breeding in Nepal were also discussed.